Reference electrodes having an extended lifetime for use in long term amperometric sensors

ABSTRACT

The present application provides Ag/AgCl based reference electrodes having an extended lifetime that are suitable for use in long term amperometric sensors. Electrochemical sensors equipped with reference electrodes described herein demonstrate considerable stability and extended lifetime in a variety of conditions.

BACKGROUND OF THE INVENTION

Enzyme-based biosensors are devices in which ananalyte-concentration-dependent biochemical reaction signal is convertedinto a measurable physical signal, such as an optical or electricalsignal. Such biosensors are widely used in the detection of analytes inclinical, environmental, agricultural and biotechnological applications.Analytes that can be measured in clinical assays of fluids of the humanbody include, for example, glucose, lactate, cholesterol, bilirubin andamino acids. The detection of analytes in biological fluids, such asblood, is important in the diagnosis and the monitoring of manydiseases.

Biosensors that detect analytes via electrical signals, such as current(amperometric biosensors) or charge (coulometric biosensors), are ofspecial interest because electron transfer is involved in thebiochemical reactions of many important bioanalytes. For example, thereaction of glucose with glucose oxidase involves electron transfer fromglucose to the enzyme to produce gluconolactone and reduced enzyme. Inan example of an amperometric glucose biosensor, glucose is oxidized byoxygen in the body fluid via a glucose oxidase-catalyzed reaction thatgenerates gluconolactone and hydrogen peroxide, then the hydrogenperoxide is electrooxidized and correlated to the concentration ofglucose in the body fluid.

Some biosensors are designed for implantation in a living animal body,such as a mammalian or a human body, merely by way of example.Typically, such biosensors have a three-electrode system provided withworking electrodes which sensitively respond to species of interest,reference electrodes which control the potentials of working electrodes,and counter electrodes which pass the electrical currents generated onthe working electrodes. Alternatively, the reference and counterelectrodes can be combined as one electrode to form a two-electrodesystem. The working electrode is typically constructed of a sensinglayer, which is in direct contact with the conductive material of theelectrode, and a diffusion-limiting membrane layer on top of the sensinglayer. The reference electrode is typically composed of Ag/AgCl, whichis fabricated via screen printing or electroplating. However, thelifetime of a screen-printed Ag/AgCl reference electrode is typicallylimited in an in vivo amperometric sensor due to dissolution of the AgClinto the surrounding tissue.

As a result, the sensor's life as a whole is often limited by the amountof Ag/AgCl available on the sensor's reference electrode. Althoughincreasing the level of Ag/AgCl loaded on the reference electrode canprolong the lifetime of the reference electrode, the small and compactsize of an implantable biosensor prevents from doing so.

Therefore, there remains a need for providing a reference electrodehaving an extended lifetime that is suitable for long term use in animplantable biosensor. The present invention addresses this need.

SUMMARY OF THE INVENTION

The present application provides Ag/AgCl based reference electrodeshaving an extended lifetime that are suitable for use in long termamperometric sensors. Electrochemical sensors equipped with referenceelectrodes described herein demonstrate considerable stability andextended lifetime in a variety of conditions.

These and other objects, advantages, and features of the invention willbecome apparent to those persons skilled in the art upon reading thedetails of the invention as more fully described below.

BRIEF DESCRIPTION OF THE DRAWINGS

The invention is best understood from the following detailed descriptionwhen read in conjunction with the accompanying drawings. It isemphasized that, according to common practice, the various features ofthe drawings are not to-scale. On the contrary, the dimensions of thevarious features are arbitrarily expanded or reduced for clarity.Included in the drawings are the following figures:

FIG. 1 shows a block diagram of an embodiment of a data monitoring andmanagement system according to the present invention;

FIG. 2 shows a block diagram of an embodiment of the transmitter unit ofthe data monitoring and management system of FIG. 1;

FIG. 3 shows a block diagram of an embodiment of the receiver/monitorunit of the data monitoring and management system of FIG. 1;

FIG. 4 shows a schematic diagram of an embodiment of an analyte sensoraccording to the present invention;

FIGS. 5A-5B show a perspective view and a cross sectional view,respectively of another embodiment an analyte sensor;

FIG. 6A is a cross-sectional schematic layout of an electrode anddielectric layers (polyester substrate (601), carbon working electrodelayer (602), dielectric layer (603), carbon conductive trace forreference electrode (604), Ag/AgCl reference electrode pad (605),dielectric layer (606), carbon counter electrode (607), and dielectriclayer (608));

FIG. 6B is a cross-sectional schematic layout of an electrode anddielectric layer covering the Ag/AgCl (polyester substrate (701), carbonworking electrode layer (702), dielectric layer (703), Ag/AgCl referenceelectrode pad (704), dielectric layer (706), carbon counter electrode(707), dielectric layer (708));

FIG. 7 shows sensor life dependence on the coverage area of Ag/AgCl ofreference electrode by dielectric;

FIG. 8 shows increase in sensor life at 66° C. as a result ofapplication of a permselective coating over Ag/AgCl;

FIG. 9 shows the AgCl dissolution rate of a standard Ag/AgCl referenceelectrode as determined by coulometric method;

FIGS. 10A-10B show regeneration of AgCl using external reference andcounter electrodes by application of electrical potential to convert Agto AgCl (FIG. 10A shows the first six hours of the experiment and FIG.10B shows the same experiment between hours 25 and 55);

FIG. 11 shows Os (III/II) potential range vs. a standard Ag/AgClreference electrode; and

FIG. 12 shows regeneration of AgCl in-situ using the Navigator™ sensor'sworking electrode as the reference, and its counter as the counterelectrode, by application of electrical potential to convert Ag to AgCl.

The figures shown herein are not necessarily drawn to scale, with somecomponents and features being exaggerated for clarity.

DETAILED DESCRIPTION OF THE INVENTION

Before the present invention is described, it is to be understood thatthis invention is not limited to particular embodiments described, assuch may, of course, vary. It is also to be understood that theterminology used herein is for the purpose of describing particularembodiments only, and is not intended to be limiting, since the scope ofthe present invention will be limited only by the appended claims.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimits of that range is also specifically disclosed. Each smaller rangebetween any stated value or intervening value in a stated range and anyother stated or intervening value in that stated range is encompassedwithin the invention. The upper and lower limits of these smaller rangesmay independently be included or excluded in the range, and each rangewhere either, neither or both limits are included in the smaller rangesis also encompassed within the invention, subject to any specificallyexcluded limit in the stated range. Where the stated range includes oneor both of the limits, ranges excluding either or both of those includedlimits are also included in the invention.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, some potential andpreferred methods and materials are now described. All publicationsmentioned herein are incorporated herein by reference to disclose anddescribe the methods and/or materials in connection with which thepublications are cited. It is understood that the present disclosuresupercedes any disclosure of an incorporated publication to the extentthere is a contradiction.

It must be noted that as used herein and in the appended claims, thesingular forms “a”, “an”, and “the” include plural referents unless thecontext clearly dictates otherwise.

The publications discussed herein are provided solely for theirdisclosure prior to the filing date of the present application. Nothingherein is to be construed as an admission that the present invention isnot entitled to antedate such publication by virtue of prior invention.Further, the dates of publication provided may be different from theactual publication dates which may need to be independently confirmed.

Reference Electrodes

In general, electrochemical sensors include a working electrode, areference electrode and a counter electrode. A working electrode is anelectrode at which the analyte, or a compound whose level depends on thelevel of the analyte, is electrooxidized or electroreduced with orwithout the agency of an electron transfer agent. A reference electroderefers to an electrode that functions as a redox electrode that providesfor measuring or controlling the potential of the working electrode. Theterm reference electrode includes both a) reference electrodes and b)reference electrodes that also function as counter electrodes (i.e.,counter/reference electrodes), unless otherwise indicated.

In order for an electrochemical sensor to function properly over anextended period of time the reference electrode must be able to maintaina stable potential over the lifetime of the measurement period. In thecase of an implanted continuous and/or automatic in vivo monitoringsystem, this period could range from hours, to day, and months, or more.The reference electrodes described herein comprise silver metal (Ag) andits silver salt (AgCl) deposited on a solid substrate. However, due tothe solubility of AgCl in an aqueous environment, a reference electrodecomprising Ag/AgCl as the reference element will be subject todissolution over an extended period of in vivo implantation.

In order to increase the period of time in which an electrochemicalsensor may remain implanted in vivo, the present invention providesreference electrodes having an extended Ag/AgCl lifetime that aresuitable for use in long term amperometric sensors. In some embodiment,a portion of the Ag/AgCl of the reference electrode is covered with animpermeable dielectric layer in order to protect the Ag/AgCl underneaththe layer from direct contact with the aqueous environment. In otherembodiment, the Ag/AgCl of the reference electrode is covered with apermselective coating that provides for low permeability of the AgCl tothe surrounding aqueous environment. In other embodiments, a briefelectric potential is applied to the Ag/AgCl reference electrode inorder to convert the silver metal (Ag) to its silver salt (AgCl) toreplenish the silver salt that has been lost due to dissolution.

Such Ag/AgCl based reference electrodes having an extended lifetime aredescribed in greater detail below.

Reference Electrode Having a Dielectric Layer

As noted above, in some embodiments, in order to increase the lifetimeof an Ag/AgCl reference electrode, a portion of the lateral surface ofthe Ag/AgCl is covered with a dielectric layer that protects the Ag/AgClunderneath from dissolution into the environment and provides areservoir of Ag/AgCl. As such, the reservoir of Ag/AgCl under thedielectric layer is capable of replacing the AgCl that is dissolved inthe aqueous environment following implantation of at least a portion ofthe reference electrode subcutaneously in a patient. By “covered” ismeant that the layer of dielectric material is disposed on the lateralsurface the Ag/AgCl, thereby covering a portion of the surface of theAg/AgCl and providing an exposed portion of the lateral surface of theAg/AgCl.

The term dielectric layer broadly refers to a thin-film structure ofdielectric material deposited on at least a portion of the Ag/AgCl of areference electrode. Most dielectric materials are solid. Examplesinclude porcelain (ceramic), mica, glass, plastics, and the oxides ofvarious metals. A suitable dielectric layer may be composed of, forexample, Al₂O₃, SiO₂, HfO₂, ZrO₂, TiO₂, La₂O₃, Y₂O₃, Gd₂O₃, GeO₂,SrTiO₃, metal silicates (e.g., Hf_(x)Si_(y)O_(z)) and/or metalaluminates (e.g., Hf_(x)Al_(y)O_(z)).

In general, the dielectric layer will cover at least a portion of theAg/AgCl sufficient to provide a reservoir of Ag/AgCl underneath thedielectric layer. The dielectric layer is disposed on at least about 5%or more of the lateral surface of the Ag/AgCl, including up to about 95%of the lateral surface of the Ag/AgCl. In some embodiments, thedielectric layer covers at least about 10%, including about 20%, about30%, about 40%, about 50%, about 60%, about 70%, about 80%, and about90%, of the lateral surface of the Ag/AgCl.

For example, as shown in FIG. 6B, in one aspect, the sensor 500 (such asthe sensor unit 101 FIG. 1), includes a substrate layer 701, and a firstconducting layer 702 such as carbon, gold, etc., disposed on at least aportion of the substrate layer 701, and which may provide the workingelectrode. Also disposed on at least a portion of the first conductinglayer 702 is a sensing layer.

A first insulation layer such as a first dielectric layer 703 isdisposed or layered on at least a portion of the first conducting layer702, and further, a Ag/AgCl conducting layer 704 may be disposed orstacked on top of at least a portion of the first insulation layer (ordielectric layer) 703.

A second insulation layer 706 such as a dielectric layer in oneembodiment may be disposed or layered on at least a portion of theAg/AgCl layer 704. Further, a third conducting layer 707 may provide thecounter electrode. It may be disposed on at least a portion of thesecond insulation layer 706. Finally, a third insulation layer 708 maybe disposed or layered on at least a portion of the third conductinglayer 707. In this manner, the sensor may be layered such that at leasta portion of each of the conducting layers is separated by a respectiveinsulation layer (for example, a dielectric layer). The embodiment ofFIG. 6B show the layers having different lengths. Some or all of thelayers may have the same or different lengths and/or widths.

Reference Electrode Having a Permaselective Coating

In some embodiments, in order to increase the lifetime of an Ag/AgClreference electrode, at lest a portion of the lateral surface of theAg/AgCl is covered with a permselective coating that protects theAg/AgCl underneath from dissolution into the environment and provides areservoir of Ag/AgCl. As such, the permselective coating prevents theAg/AgCl underneath to dissolve into the aqueous environment, therebyproviding a constant level of Ag/AgCl to maintain a stable potentialover the lifetime of the measurement period, such as followingimplantation of at least a portion of the reference electrodesubcutaneously in a patient.

In some embodiments the permselective coating is a copolymer ofpolyvinyl pyridine and styrene. In such embodiments, the polyvinylpyridine will be loaded with at 5% styrene, including about 10%, about12%, about 15%, about 17%, and about 20% or more. In some embodiments,the permselective layer comprises polyvinyl pyridine with about a 10%loading of styrene. Suitable copolymers include,poly(2-vinylpyridine-co-styrene), poly(4-vinylpyridine-co-styrene), andthe like.

Such a permselective layer for use in limiting the dissolution of theAg/AgCl of the reference electrode into the aqueous environment may bedifferent that a flux limiting membrane applied over the sensing layerof the working electrode that limits the flux of analytes to the enzymespresent in the sensing layer. As such, in some embodiments, theelectrode will further comprise a second flux limiting membrane.

In Situ Renewal of Ag/AgCl of a Reference Electrode

In some embodiments level of AgCl on a reference electrode of aelectrochemical sensor is replenished by applying an electricalpotential across the reference electrode and another electrode for aperiod of time sufficient to convert Ag to AgCl in order to replenishthe level of AgCl present on the reference electrode. In general,following implantation of at least a portion of the reference electrodesubcutaneously in a patient, a portion of the AgCl present on thereference electrode will dissolve into the aqueous environment. As aresult of the loss of AgCl, the electrochemical sensor will not be ableto maintain a stable potential over the lifetime of the measurementperiod. An electrical potential applied to the reference electrode willresult in the oxidation of the silver metal (Ag) portion of to provideAg⁺. The Ag⁺, will then combine with Cl⁻ present in the in vivoenvironment to form silver salt (AgCl) on the reference electrode,thereby replenishing the level of AgCl present on the referenceelectrode.

The electrical potential applied to the reference electrode must be at asufficient level and for a sufficient period of time to provide forconversion of a sufficient level of Ag to Ag⁺. The electrical potentialapplied will include at least about +50 mV, such as about +75 mV, about+100 mV, +125 mV, +150 mV, +175 mV, +200 mV, +250 mV, etc. In addition,the electrical potential is applied for a duration of at least 30seconds, including about 45 seconds, about 1 minute, about 2 minutes,about three minutes or more. It will be appreciated by one of skill inthe art that the level as well the duration of the electrical potentialapplied may be adjusted to provide a suitable amount of Ag conversion.For example, a low level electrical potential, such as +50 mV, withlonger duration, such as 2 minutes. As the level of electrical potentialis increased, the duration at which the electrical potential is appliedmay be decreased.

The application of the electrical potential can also be repeated one ormore times during the period in which a portion of the referenceelectrode is placed subcutaneously in a patient. This period may lastfrom about 1 day to about 3 days, about 5 days, about 1 week, about 2weeks, about 3, weeks, about 1 month, about 2 months or more. As such,the application of the electrical potential may be repeated about 2times, about 3 times, about 4 times, about 5 times, about 6 times, about7 times, about 8 times, about 9 times, about 10 times, or more. It willbe appreciated by one skilled in the art that the number of times anelectrical potential may be applied to the reference electrode in orderto replenish the level of AgCl present on the reference electrode willbe limited by the level of Ag present on the reference electrode that isavailable for conversion to AgCl.

Electrochemical Sensors

Generally, embodiments of the present invention relate to methods anddevices for detecting at least one analyte, such as glucose, in bodyfluid. Embodiments relate to the continuous and/or automatic in vivomonitoring of the level of one or more analytes using a continuousanalyte monitoring system that includes an analyte sensor at least aportion of which is to be positioned beneath a skin surface of a userfor a period of time and/or the discrete monitoring of one or moreanalytes using an in vitro blood glucose (“BG”) meter and an analytetest strip. Embodiments include combined or combinedable devices,systems and methods and/or transferring data between an in vivocontinuous system and a BG meter system.

An electrochemical sensor that includes at least one Ag/AgCl referenceelectrode having an extended lifetime can be formed on a substrate. Thesensor may also include at least one counter electrode (orcounter/reference electrode) and/or at least one reference electrode. An“electrochemical sensor” is a device configured to detect the presenceand/or measure the level of an analyte in a sample, via anelectrochemical oxidation or reduction reaction on the sensor, or via asequence of chemical reactions where at least one of the chemicalreactions is an electrochemical oxidation or reduction reactions on thesensor. These reactions are transduced to an electrical signal that canbe correlated to an amount, concentration, or level of an analyte in thesample.

Accordingly, embodiments include analyte monitoring devices and systemsthat include an analyte sensor—at least a portion of which ispositionable beneath the skin of the user—for the in vivo detection, ofan analyte, such as glucose, lactate, and the like, in a body fluid.Embodiments include wholly implantable analyte sensors and analytesensors in which only a portion of the sensor is positioned under theskin and a portion of the sensor resides above the skin, e.g., forcontact to a transmitter, receiver, transceiver, processor, etc. Thesensor may be, for example, subcutaneously positionable in a patient forthe continuous or periodic monitoring of a level of an analyte in apatient's interstitial fluid. For the purposes of this description,continuous monitoring and periodic monitoring will be usedinterchangeably, unless noted otherwise. The sensor response may becorrelated and/or converted to analyte levels in blood or other fluids.In certain embodiments, an analyte sensor may be positioned in contactwith interstitial fluid to detect the level of glucose, which detectedglucose may be used to infer the glucose level in the patient'sbloodstream. Analyte sensors may be insertable into a vein, artery, orother portion of the body containing fluid. Embodiments of the analytesensors of the subject invention having an Ag/AgCl reference electrodehaving an extended lifetime may be configured for monitoring the levelof the analyte over a time period which may range from minutes, hours,days, weeks, to months, or longer.

Of interest are analyte sensors, such as glucose sensors, having anAg/AgCl reference electrode having an extended lifetime, that arecapable of in vivo detection of an analyte for about one hour or more,e.g., about a few hours or more, e.g., about a few days of more, e.g.,about three or more days, e.g., about five days or more, e.g., aboutseven days or more, e.g., about several weeks or at least one month ormore. Future analyte levels may be predicted based on informationobtained, e.g., the current analyte level at time t₀, the rate of changeof the analyte, etc. Predictive alarms may notify the user of apredicted analyte levels that may be of concern in advance of the user'sanalyte level reaching the future level. This provides the user anopportunity to take corrective action.

FIG. 1 shows a data monitoring and management system such as, forexample, an analyte (e.g., glucose) monitoring system 100 in accordancewith certain embodiments. Embodiments of the subject invention arefurther described primarily with respect to glucose monitoring devicesand systems, and methods of glucose detection, for convenience only andsuch description is in no way intended to limit the scope of theinvention. It is to be understood that the analyte monitoring system maybe configured to monitor a variety of analytes at the same time or atdifferent times.

Analytes that may be monitored include, but are not limited to, acetylcholine, amylase, bilirubin, cholesterol, chorionic gonadotropin,creatine kinase (e.g., CK-MB), creatine, creatinine, DNA, fructosamine,glucose, glutamine, growth hormones, hormones, ketone bodies, lactate,peroxide, prostate-specific antigen, prothrombin, RNA, thyroidstimulating hormone, and troponin. The concentration of drugs, such as,for example, antibiotics (e.g., gentamicin, vancomycin, and the like),digitoxin, digoxin, drugs of abuse, theophylline, and warfarin, may alsobe monitored. In those embodiments that monitor more than one analyte,the analytes may be monitored at the same or different times.

The analyte monitoring system 100 includes a sensor 101, a dataprocessing unit 102 connectable to the sensor 101, and a primaryreceiver unit 104 which is configured to communicate with the dataprocessing unit 102 via a communication link 103. In certainembodiments, the primary receiver unit 104 may be further configured totransmit data to a data processing terminal 105 to evaluate or otherwiseprocess or format data received by the primary receiver unit 104. Thedata processing terminal 105 may be configured to receive data directlyfrom the data processing unit 102 via a communication link which mayoptionally be configured for bi-directional communication. Further, thedata processing unit 102 may include a transmitter or a transceiver totransmit and/or receive data to and/or from the primary receiver unit104 and/or the data processing terminal 105 and/or optionally thesecondary receiver unit 106.

Also shown in FIG. 1 is an optional secondary receiver unit 106 which isoperatively coupled to the communication link and configured to receivedata transmitted from the data processing unit 102. The secondaryreceiver unit 106 may be configured to communicate with the primaryreceiver unit 104, as well as the data processing terminal 105. Thesecondary receiver unit 106 may be configured for bi-directionalwireless communication with each of the primary receiver unit 104 andthe data processing terminal 105. As discussed in further detail below,in certain embodiments the secondary receiver unit 106 may be ade-featured receiver as compared to the primary receiver, i.e., thesecondary receiver may include a limited or minimal number of functionsand features as compared with the primary receiver unit 104. As such,the secondary receiver unit 106 may include a smaller (in one or more,including all, dimensions), compact housing or embodied in a device suchas a wrist watch, arm band, etc., for example. Alternatively, thesecondary receiver unit 106 may be configured with the same orsubstantially similar functions and features as the primary receiverunit 104. The secondary receiver unit 106 may include a docking portionto be mated with a docking cradle unit for placement by, e.g., thebedside for night time monitoring, and/or a bi-directional communicationdevice. A docking cradle may recharge a powers supply.

Only one sensor 101, data processing unit 102 and data processingterminal 105 are shown in the embodiment of the analyte monitoringsystem 100 illustrated in FIG. 1. However, it will be appreciated by oneof ordinary skill in the art that the analyte monitoring system 100 mayinclude more than one sensor 101 and/or more than one data processingunit 102, and/or more than one data processing terminal 105. Multiplesensors may be positioned in a patient for analyte monitoring at thesame or different times. In certain embodiments, analyte informationobtained by a first positioned sensor may be employed as a comparison toanalyte information obtained by a second sensor. This may be useful toconfirm or validate analyte information obtained from one or both of thesensors. Such redundancy may be useful if analyte information iscontemplated in critical therapy-related decisions. In certainembodiments, a first sensor may be used to calibrate a second sensor.

The analyte monitoring system 100 may be a continuous monitoring system,or semi-continuous, or a discrete monitoring system. In amulti-component environment, each component may be configured to beuniquely identified by one or more of the other components in the systemso that communication conflict may be readily resolved between thevarious components within the analyte monitoring system 100. Forexample, unique IDs, communication channels, and the like, may be used.

In certain embodiments, the sensor 101 is physically positioned in or onthe body of a user whose analyte level is being monitored. The sensor101 may be configured to at least periodically sample the analyte levelof the user and convert the sampled analyte level into a correspondingsignal for transmission by the data processing unit 102. The dataprocessing unit 102 is coupleable to the sensor 101 so that both devicesare positioned in or on the user's body, with at least a portion of theanalyte sensor 101 positioned transcutaneously. The data processing unitmay include a fixation element such as adhesive or the like to secure itto the user's body. A mount (not shown) attachable to the user andmateable with the unit 102 may be used. For example, a mount may includean adhesive surface. The data processing unit 102 performs dataprocessing functions, where such functions may include but are notlimited to, filtering and encoding of data signals, each of whichcorresponds to a sampled analyte level of the user, for transmission tothe primary receiver unit 104 via the communication link 103. In oneembodiment, the sensor 101 or the data processing unit 102 or a combinedsensor/data processing unit may be wholly implantable under the skinlayer of the user.

In certain embodiments, the primary receiver unit 104 may include ananalog interface section including and RF receiver and an antenna thatis configured to communicate with the data processing unit 102 via thecommunication link 103, and a data processing section for processing thereceived data from the data processing unit 102 such as data decoding,error detection and correction, data clock generation, data bitrecovery, etc., or any combination thereof.

In operation, the primary receiver unit 104 in certain embodiments isconfigured to synchronize with the data processing unit 102 to uniquelyidentify the data processing unit 102, based on, for example, anidentification information of the data processing unit 102, andthereafter, to periodically receive signals transmitted from the dataprocessing unit 102 associated with the monitored analyte levelsdetected by the sensor 101.

Referring again to FIG. 1, the data processing terminal 105 may includea personal computer, a portable computer such as a laptop or a handhelddevice (e.g., personal digital assistants (PDAs), telephone such as acellular phone (e.g., a multimedia and Internet-enabled mobile phonesuch as an iPhone™ or similar phone), mp3 player, pager, and the like),drug delivery device, each of which may be configured for datacommunication with the receiver via a wired or a wireless connection.Additionally, the data processing terminal 105 may further be connectedto a data network (not shown) for storing, retrieving, updating, and/oranalyzing data corresponding to the detected analyte level of the user.

The data processing terminal 105 may include an infusion device such asan insulin infusion pump or the like, which may be configured toadminister insulin to patients, and which may be configured tocommunicate with the primary receiver unit 104 for receiving, amongothers, the measured analyte level. Alternatively, the primary receiverunit 104 may be configured to integrate an infusion device therein sothat the primary receiver unit 104 is configured to administer insulin(or other appropriate drug) therapy to patients, for example, foradministering and modifying basal profiles, as well as for determiningappropriate boluses for administration based on, among others, thedetected analyte levels received from the data processing unit 102. Aninfusion device may be an external device or an internal device (whollyimplantable in a user).

In certain embodiments, the data processing terminal 105, which mayinclude an insulin pump, may be configured to receive the analytesignals from the data processing unit 102, and thus, incorporate thefunctions of the primary receiver unit 104 including data processing formanaging the patient's insulin therapy and analyte monitoring. Incertain embodiments, the communication link 103 as well as one or moreof the other communication interfaces shown in FIG. 1, may use one ormore of: an RF communication protocol, an infrared communicationprotocol, a Bluetooth enabled communication protocol, an 802.11xwireless communication protocol, or an equivalent wireless communicationprotocol which would allow secure, wireless communication of severalunits (for example, per HIPPA requirements), while avoiding potentialdata collision and interference.

FIG. 2 shows a block diagram of an embodiment of a data processing unitof the data monitoring and detection system shown in FIG. 1. User inputand/or interface components may be included or a data processing unitmay be free of user input and/or interface components. In certainembodiments, one or more application-specific integrated circuits (ASIC)may be used to implement one or more functions or routins associatedwith the operations of the data processing unit (and/or receiver unit)using for example one or more state machines and buffers.

As can be seen in the embodiment of FIG. 2, the sensor unit 101 (FIG. 1)includes four contacts, three of which are electrodes—work electrode (W)210, reference electrode (R) 212, and counter electrode (C) 213, eachoperatively coupled to the analog interface 201 of the data processingunit 102. This embodiment also shows optional guard contact (G) 211.Fewer or greater electrodes may be employed. For example, the counterand reference electrode functions may be served by a singlecounter/reference electrode, there may be more than one workingelectrode and/or reference electrode and/or counter electrode, etc.

FIG. 3 is a block diagram of an embodiment of a receiver/monitor unitsuch as the primary receiver unit 104 of the data monitoring andmanagement system shown in FIG. 1. The primary receiver unit 104includes one or more of: a blood glucose test strip interface 301, an RFreceiver 302, an input 303, a temperature detection section 304, and aclock 305, each of which is operatively coupled to a processing andstorage section 307. The primary receiver unit 104 also includes a powersupply 306 operatively coupled to a power conversion and monitoringsection 308. Further, the power conversion and monitoring section 308 isalso coupled to the receiver processor 307. Moreover, also shown are areceiver serial communication section 309, and an output 310, eachoperatively coupled to the processing and storage unit 307. The receivermay include user input and/or interface components or may be free ofuser input and/or interface components.

In certain embodiments, the test strip interface 301 includes a glucoselevel testing portion to receive a blood (or other body fluid sample)glucose test or information related thereto. For example, the interfacemay include a test strip port to receive a glucose test strip. Thedevice may determine the glucose level of the test strip, and optionallydisplay (or otherwise notice) the glucose level on the output 310 of theprimary receiver unit 104. Any suitable test strip may be employed,e.g., test strips that only require a very small amount (e.g., onemicroliter or less, e.g., 0.5 microliter or less, e.g., 0.1 microliteror less), of applied sample to the strip in order to obtain accurateglucose information, e.g. FreeStyle® blood glucose test strips fromAbbott Diabetes Care, Inc. Glucose information obtained by the in vitroglucose testing device may be used for a variety of purposes,computations, etc. For example, the information may be used to calibratesensor 101, confirm results of the sensor 101 to increase the confidencethereof (e.g., in instances in which information obtained by sensor 101is employed in therapy related decisions), etc.

In further embodiments, the data processing unit 102 and/or the primaryreceiver unit 104 and/or the secondary receiver unit 105, and/or thedata processing terminal/infusion section 105 may be configured toreceive the blood glucose value wirelessly over a communication linkfrom, for example, a blood glucose meter. In further embodiments, a usermanipulating or using the analyte monitoring system 100 (FIG. 1) maymanually input the blood glucose value using, for example, a userinterface (for example, a keyboard, keypad, voice commands, and thelike) incorporated in the one or more of the data processing unit 102,the primary receiver unit 104, secondary receiver unit 105, or the dataprocessing terminal/infusion section 105.

Additional detailed descriptions are provided in U.S. Pat. Nos.5,262,035; 5,264,104; 5,262,305; 5,320,715; 5,593,852; 6,175,752;6,650,471; 6,746,582, and in application Ser. No. 10/745,878 filed Dec.26, 2003 entitled “Continuous Glucose Monitoring System and Methods ofUse”, each of which is incorporated herein by reference.

FIG. 4 schematically shows an embodiment of an analyte sensor inaccordance with the present invention. This sensor embodiment includeselectrodes 401, 402 and 403 on a base 404. Electrodes (and/or otherfeatures) may be applied or otherwise processed using any suitabletechnology, e.g., chemical vapor deposition (CVD), physical vapordeposition, sputtering, reactive sputtering, printing, coating, ablating(e.g., laser ablation), painting, dip coating, etching, and the like.Materials include but are not limited to aluminum, carbon (such asgraphite), cobalt, copper, gallium, gold, indium, iridium, iron, lead,magnesium, mercury (as an amalgam), nickel, niobium, osmium, palladium,platinum, rhenium, rhodium, selenium, silicon (e.g., dopedpolycrystalline silicon), silver, tantalum, tin, titanium, tungsten,uranium, vanadium, zinc, zirconium, mixtures thereof, and alloys,oxides, or metallic compounds of these elements.

The sensor may be wholly implantable in a user or may be configured sothat only a portion is positioned within (internal) a user and anotherportion outside (external) a user. For example, the sensor 400 mayinclude a portion positionable above a surface of the skin 410, and aportion positioned below the skin. In such embodiments, the externalportion may include contacts (connected to respective electrodes of thesecond portion by traces) to connect to another device also external tothe user such as a transmitter unit. While the embodiment of FIG. 4shows three electrodes side-by-side on the same surface of base 404,other configurations are contemplated, e.g., fewer or greaterelectrodes, some or all electrodes on different surfaces of the base orpresent on another base, some or all electrodes stacked together,electrodes of differing materials and dimensions, etc.

FIG. 5A shows a perspective view of an embodiment of an electrochemicalanalyte sensor 500 having a first portion (which in this embodiment maybe characterized as a major portion) positionable above a surface of theskin 510, and a second portion (which in this embodiment may becharacterized as a minor portion) that includes an insertion tip 530positionable below the skin, e.g., penetrating through the skin andinto, e.g., the subcutaneous space 520, in contact with the user'sbiofluid such as interstitial fluid. Contact portions of a workingelectrode 501, a reference electrode 502, and a counter electrode 503are positioned on the portion of the sensor 500 situated above the skinsurface 510. Working electrode 501, a reference electrode 502, and acounter electrode 503 are shown at the second section and particularlyat the insertion tip 530. Traces may be provided from the electrode atthe tip to the contact, as shown in FIG. 5A. It is to be understood thatgreater or fewer electrodes may be provided on a sensor. For example, asensor may include more than one working electrode and/or the counterand reference electrodes may be a single counter/reference electrode,etc.

FIG. 5B shows a cross sectional view of a portion of the sensor 500 ofFIG. 5A. The electrodes 501, 502 and 503, of the sensor 500 as well asthe substrate and the dielectric layers are provided in a layeredconfiguration or construction. For example, as shown in FIG. 5B, in oneaspect, the sensor 500 (such as the sensor unit 101 FIG. 1), includes asubstrate layer 504, and a first conducting layer 501 such as carbon,gold, etc., disposed on at least a portion of the substrate layer 504,and which may provide the working electrode. Also shown disposed on atleast a portion of the first conducting layer 501 is a sensing layer508.

A first insulation layer such as a first dielectric layer 505 isdisposed or layered on at least a portion of the first conducting layer501, and further, a second conducting layer 509 may be disposed orstacked on top of at least a portion of the first insulation layer (ordielectric layer) 505. As shown in FIG. 5B, the second conducting layer509 may provide the reference electrode 502, as described herein havingan extended lifetime, which includes a layer of silver/silver chloride(Ag/AgCl).

A second insulation layer 506 such as a dielectric layer in oneembodiment may be disposed or layered on at least a portion of thesecond conducting layer 509. Further, a third conducting layer 503 mayprovide the counter electrode 503. It may be disposed on at least aportion of the second insulation layer 506. Finally, a third insulationlayer may be disposed or layered on at least a portion of the thirdconducting layer 503. In this manner, the sensor 500 may be layered suchthat at least a portion of each of the conducting layers is separated bya respective insulation layer (for example, a dielectric layer). Theembodiment of FIGS. 5A and 5B show the layers having different lengths.Some or all of the layers may have the same or different lengths and/orwidths.

In certain embodiments, some or all of the electrodes 501, 502, 503 maybe provided on the same side of the substrate 504 in the layeredconstruction as described above, or alternatively, may be provided in aco-planar manner such that two or more electrodes may be positioned onthe same plane (e.g., side-by side (e.g., parallel) or angled relativeto each other) on the substrate 504. For example, co-planar electrodesmay include a suitable spacing there between and/or include dielectricmaterial or insulation material disposed between the conductinglayers/electrodes. Furthermore, in certain embodiments one or more ofthe electrodes 501, 502, 503 may be disposed on opposing sides of thesubstrate 504. In such embodiments, contact pads may be one the same ordifferent sides of the substrate. For example, an electrode may be on afirst side and its respective contact may be on a second side, e.g., atrace connecting the electrode and the contact may traverse through thesubstrate.

As noted above, analyte sensors may include an analyte-responsive enzymeto provide a sensing component or sensing layer. Some analytes, such asoxygen, can be directly electrooxidized or electroreduced on a sensor,and more specifically at least on a working electrode of a sensor. Otheranalytes, such as glucose and lactate, require the presence of at leastone electron transfer agent and/or at least one catalyst to facilitatethe electrooxidation or electroreduction of the analyte. Catalysts mayalso be used for those analyte, such as oxygen, that can be directlyelectrooxidized or electroreduced on the working electrode. For theseanalytes, each working electrode includes a sensing layer (see forexample sensing layer 408 of FIG. 5B) proximate to or on a surface of aworking electrode. In many embodiments, a sensing layer is formed nearor on only a small portion of at least a working electrode.

The sensing layer includes one or more components designed to facilitatethe electrochemical oxidation or reduction of the analyte. The sensinglayer may include, for example, a catalyst to catalyze a reaction of theanalyte and produce a response at the working electrode, an electrontransfer agent to transfer electrons between the analyte and the workingelectrode (or other component), or both.

A variety of different sensing layer configurations may be used. Incertain embodiments, the sensing layer is deposited on the conductivematerial of a working electrode. The sensing layer may extend beyond theconductive material of the working electrode. In some cases, the sensinglayer may also extend over other electrodes, e.g., over the counterelectrode and/or reference electrode (or counter/reference is provided).

A sensing layer that is in direct contact with the working electrode maycontain an electron transfer agent to transfer electrons directly orindirectly between the analyte and the working electrode, and/or acatalyst to facilitate a reaction of the analyte. For example, aglucose, lactate, or oxygen electrode may be formed having a sensinglayer which contains a catalyst, such as glucose oxidase, lactateoxidase, or laccase, respectively, and an electron transfer agent thatfacilitates the electrooxidation of the glucose, lactate, or oxygen,respectively.

In other embodiments the sensing layer is not deposited directly on theworking electrode. Instead, the sensing layer 64 may be spaced apartfrom the working electrode, and separated from the working electrode,e.g., by a separation layer. A separation layer may include one or moremembranes or films or a physical distance. In addition to separating theworking electrode from the sensing layer the separation layer may alsoact as a mass transport limiting layer and/or an interferent eliminatinglayer and/or a biocompatible layer.

In certain embodiments which include more than one working electrode,one or more of the working electrodes may not have a correspondingsensing layer, or may have a sensing layer which does not contain one ormore components (e.g., an electron transfer agent and/or catalyst)needed to electrolyze the analyte. Thus, the signal at this workingelectrode may correspond to background signal which may be removed fromthe analyte signal obtained from one or more other working electrodesthat are associated with fully-functional sensing layers by, forexample, subtracting the signal.

In certain embodiments, the sensing layer includes one or more electrontransfer agents. Electron transfer agents that may be employed areelectroreducible and electrooxidizable ions or molecules having redoxpotentials that are a few hundred millivolts above or below the redoxpotential of the standard calomel electrode (SCE). The electron transferagent may be organic, organometallic, or inorganic. Examples of organicredox species are quinones and species that in their oxidized state havequinoid structures, such as Nile blue and indophenol. Examples oforganometallic redox species are metallocenes such as ferrocene.Examples of inorganic redox species are hexacyanoferrate (III),ruthenium hexamine etc.

In certain embodiments, electron transfer agents have structures orcharges which prevent or substantially reduce the diffusional loss ofthe electron transfer agent during the period of time that the sample isbeing analyzed. For example, electron transfer agents include but arenot limited to a redox species, e.g., bound to a polymer which can inturn be disposed on or near the working electrode. The bond between theredox species and the polymer may be covalent, coordinative, or ionic.Although any organic, organometallic or inorganic redox species may bebound to a polymer and used as an electron transfer agent, in certainembodiments the redox species is a transition metal compound or complex,e.g., osmium, ruthenium, iron, and cobalt compounds or complexes. Itwill be recognized that many redox species described for use with apolymeric component may also be used, without a polymeric component.

One type of polymeric electron transfer agent contains a redox speciescovalently bound in a polymeric composition. An example of this type ofmediator is poly(vinylferrocene). Another type of electron transferagent contains an ionically-bound redox species. This type of mediatormay include a charged polymer coupled to an oppositely charged redoxspecies. Examples of this type of mediator include a negatively chargedpolymer coupled to a positively charged redox species such as an osmiumor ruthenium polypyridyl cation. Another example of an ionically-boundmediator is a positively charged polymer such as quaternizedpoly(4-vinyl pyridine) or poly(1-vinyl imidazole) coupled to anegatively charged redox species such as ferricyanide or ferrocyanide.In other embodiments, electron transfer agents include a redox speciescoordinatively bound to a polymer. For example, the mediator may beformed by coordination of an osmium or cobalt 2,2′-bipyridyl complex topoly(1-vinyl imidazole) or poly(4-vinyl pyridine).

Suitable electron transfer agents are osmium transition metal complexeswith one or more ligands, each ligand having a nitrogen-containingheterocycle such as 2,2′-bipyridine, 1,10-phenanthroline, 1-methyl,2-pyridyl biimidazole, or derivatives thereof. The electron transferagents may also have one or more ligands covalently bound in a polymer,each ligand having at least one nitrogen-containing heterocycle, such aspyridine, imidazole, or derivatives thereof. One example of an electrontransfer agent includes (a) a polymer or copolymer having pyridine orimidazole functional groups and (b) osmium cations complexed with twoligands, each ligand containing 2,2′-bipyridine, 1,10-phenanthroline, orderivatives thereof, the two ligands not necessarily being the same.Some derivatives of 2,2′-bipyridine for complexation with the osmiumcation include but are not limited to 4,4′-dimethyl-2,2′-bipyridine andmono-, di-, and polyalkoxy-2,2′-bipyridines, such as4,4′-dimethoxy-2,2′-bipyridine. Derivatives of 1,10-phenanthroline forcomplexation with the osmium cation include but are not limited to4,7-dimethyl-1,10-phenanthroline and mono, di-, andpolyalkoxy-1,10-phenanthrolines, such as4,7-dimethoxy-1,10-phenanthroline. Polymers for complexation with theosmium cation include but are not limited to polymers and copolymers ofpoly(1-vinyl imidazole) (referred to as “PVI”) and poly(4-vinylpyridine) (referred to as “PVP”). Suitable copolymer substituents ofpoly(1-vinyl imidazole) include acrylonitrile, acrylamide, andsubstituted or quaternized N-vinyl imidazole, e.g., electron transferagents with osmium complexed to a polymer or copolymer of poly(1-vinylimidazole).

Embodiments may employ electron transfer agents having a redox potentialranging from about −200 mV to about +200 mV versus the standard calomelelectrode (SCE). The sensing layer may also include a catalyst which iscapable of catalyzing a reaction of the analyte. The catalyst may also,in some embodiments, act as an electron transfer agent. One example of asuitable catalyst is an enzyme which catalyzes a reaction of theanalyte. For example, a catalyst, such as a glucose oxidase, glucosedehydrogenase (e.g., pyrroloquinoline quinone (PQQ), dependent glucosedehydrogenase, flavine adenine dinucleotide (FAD) dependent glucosedehydrogenase, or nicotinamide adenine dinucleotide (NAD) dependentglucose dehydrogenase), may be used when the analyte of interest isglucose. A lactate oxidase or lactate dehydrogenase may be used when theanalyte of interest is lactate. Laccase may be used when the analyte ofinterest is oxygen or when oxygen is generated or consumed in responseto a reaction of the analyte.

The sensing layer may also include a catalyst which is capable ofcatalyzing a reaction of the analyte. The catalyst may also, in someembodiments, act as an electron transfer agent. One example of asuitable catalyst is an enzyme which catalyzes a reaction of theanalyte. For example, a catalyst, such as a glucose oxidase, glucosedehydrogenase (e.g., pyrroloquinoline quinone (PQQ), dependent glucosedehydrogenase or oligosaccharide dehydrogenase, flavine adeninedinucleotide (FAD) dependent glucose dehydrogenase, nicotinamide adeninedinucleotide (NAD) dependent glucose dehydrogenase), may be used whenthe analyte of interest is glucose. A lactate oxidase or lactatedehydrogenase may be used when the analyte of interest is lactate.Laccase may be used when the analyte of interest is oxygen or whenoxygen is generated or consumed in response to a reaction of theanalyte.

In certain embodiments, a catalyst may be attached to a polymer, crosslinking the catalyst with another electron transfer agent (which, asdescribed above, may be polymeric. A second catalyst may also be used incertain embodiments. This second catalyst may be used to catalyze areaction of a product compound resulting from the catalyzed reaction ofthe analyte. The second catalyst may operate with an electron transferagent to electrolyze the product compound to generate a signal at theworking electrode. Alternatively, a second catalyst may be provided inan interferent-eliminating layer to catalyze reactions that removeinterferents.

Certain embodiments include a Wired Enzyme™ sensing layer (AbbottDiabetes Care) that works at a gentle oxidizing potential, e.g., apotential of about +40 mV vs. Ag/AgCl. This sensing layer uses an osmium(Os)-based mediator designed for low potential operation and is stablyanchored in a polymeric layer. Accordingly, in certain embodiments thesensing element is redox active component that includes (1) Osmium-basedmediator molecules attached by stable (bidente) ligands anchored to apolymeric backbone, and (2) glucose oxidase enzyme molecules. These twoconstituents are crosslinked together.

A mass transport limiting layer (not shown), e.g., an analyte fluxmodulating layer, may be included with the sensor to act as adiffusion-limiting barrier to reduce the rate of mass transport of theanalyte, for example, glucose or lactate, into the region around theworking electrodes. The mass transport limiting layers are useful inlimiting the flux of an analyte to a working electrode in anelectrochemical sensor so that the sensor is linearly responsive over alarge range of analyte concentrations and is easily calibrated. Masstransport limiting layers may include polymers and may be biocompatible.A mass transport limiting layer may provide many functions, e.g.,biocompatibility and/or interferent-eliminating, etc.

In certain embodiments, a mass transport limiting layer is a membranecomposed of crosslinked polymers containing heterocyclic nitrogengroups, such as polymers of polyvinylpyridine and polyvinylimidazole.Embodiments also include membranes that are made of a polyurethane, orpolyether urethane, or chemically related material, or membranes thatare made of silicone, and the like.

A membrane may be formed by crosslinking in situ a polymer, modifiedwith a zwitterionic moiety, a non-pyridine copolymer component, andoptionally another moiety that is either hydrophilic or hydrophobic,and/or has other desirable properties, in an alcohol-buffer solution.The modified polymer may be made from a precursor polymer containingheterocyclic nitrogen groups. For example, a precursor polymer may bepolyvinylpyridine or polyvinylimidazole. Optionally, hydrophilic orhydrophobic modifiers may be used to “fine-tune” the permeability of theresulting membrane to an analyte of interest. Optional hydrophilicmodifiers, such as poly(ethylene glycol), hydroxyl or polyhydroxylmodifiers, may be used to enhance the biocompatibility of the polymer orthe resulting membrane.

A membrane may be formed in situ by applying an alcohol-buffer solutionof a crosslinker and a modified polymer over an enzyme-containingsensing layer and allowing the solution to cure for about one to twodays or other appropriate time period. The crosslinker-polymer solutionmay be applied to the sensing layer by placing a droplet or droplets ofthe solution on the sensor, by dipping the sensor into the solution, orthe like. Generally, the thickness of the membrane is controlled by theconcentration of the solution, by the number of droplets of the solutionapplied, by the number of times the sensor is dipped in the solution, orby any combination of these factors. A membrane applied in this mannermay have any combination of the following functions: (1) mass transportlimitation, i.e., reduction of the flux of analyte that can reach thesensing layer, (2) biocompatibility enhancement, or (3) interferentreduction.

In certain embodiments, the sensing system detects hydrogen peroxide toinfer glucose levels. For example, a hydrogen peroxide-detecting sensormay be constructed in which a sensing layer includes enzyme such asglucose oxides, glucose dehydrogensae, or the like, and is positionedproximate to the working electrode. The sensing layer may be covered byone or more layers, e.g., a membrane that is selectively permeable toglucose. Once the glucose passes through the membrane, it is oxidized bythe enzyme and reduced glucose oxidase can then be oxidized by reactingwith molecular oxygen to produce hydrogen peroxide.

Certain embodiments include a hydrogen peroxide-detecting sensorconstructed from a sensing layer prepared by crosslinking two componentstogether, for example: (1) a redox compound such as a redox polymercontaining pendent Os polypyridyl complexes with oxidation potentials ofabout +200 mV vs. SCE, and (2) periodate oxidized horseradish peroxidase(HRP). Such a sensor functions in a reductive mode; the workingelectrode is controlled at a potential negative to that of the Oscomplex, resulting in mediated reduction of hydrogen peroxide throughthe HRP catalyst.

In another example, a potentiometric sensor can be constructed asfollows. A glucose-sensing layer is constructed by crosslinking together(1) a redox polymer containing pendent Os polypyridyl complexes withoxidation potentials from about −200 mV to +200 mV vs. SCE, and (2)glucose oxidase. This sensor can then be used in a potentiometric mode,by exposing the sensor to a glucose containing solution, underconditions of zero current flow, and allowing the ratio ofreduced/oxidized Os to reach an equilibrium value. The reduced/oxidizedOs ratio varies in a reproducible way with the glucose concentration,and will cause the electrode's potential to vary in a similar way.

The substrate may be formed using a variety of non-conducting materials,including, for example, polymeric or plastic materials and ceramicmaterials. Suitable materials for a particular sensor may be determined,at least in part, based on the desired use of the sensor and propertiesof the materials.

In some embodiments, the substrate is flexible. For example, if thesensor is configured for implantation into a patient, then the sensormay be made flexible (although rigid sensors may also be used forimplantable sensors) to reduce pain to the patient and damage to thetissue caused by the implantation of and/or the wearing of the sensor. Aflexible substrate often increases the patient's comfort and allows awider range of activities. Suitable materials for a flexible substrateinclude, for example, non-conducting plastic or polymeric materials andother non-conducting, flexible, deformable materials. Examples of usefulplastic or polymeric materials include thermoplastics such aspolycarbonates, polyesters (e.g., Mylar™ and polyethylene terephthalate(PET)), polyvinyl chloride (PVC), polyurethanes, polyethers, polyamides,polyimides, or copolymers of these thermoplastics, such as PETG(glycol-modified polyethylene terephthalate).

In other embodiments, the sensors are made using a relatively rigidsubstrate to, for example, provide structural support against bending orbreaking. Examples of rigid materials that may be used as the substrateinclude poorly conducting ceramics, such as aluminum oxide and silicondioxide. One advantage of an implantable sensor having a rigid substrateis that the sensor may have a sharp point and/or a sharp edge to aid inimplantation of a sensor without an additional insertion device.

It will be appreciated that for many sensors and sensor applications,both rigid and flexible sensors will operate adequately. The flexibilityof the sensor may also be controlled and varied along a continuum bychanging, for example, the composition and/or thickness of thesubstrate.

In addition to considerations regarding flexibility, it is oftendesirable that implantable sensors should have a substrate which isphysiologically harmless, for example, a substrate approved by aregulatory agency or private institution for in vivo use.

The sensor may include optional features to facilitate insertion of animplantable sensor. For example, the sensor may be pointed at the tip toease insertion. In addition, the sensor may include a barb which assistsin anchoring the sensor within the tissue of the patient duringoperation of the sensor. However, the barb is typically small enough sothat little damage is caused to the subcutaneous tissue when the sensoris removed for replacement.

An implantable sensor may also, optionally, have an anticlotting agentdisposed on a portion the substrate which is implanted into a patient.This anticlotting agent may reduce or eliminate the clotting of blood orother body fluid around the sensor, particularly after insertion of thesensor. Blood clots may foul the sensor or irreproducibly reduce theamount of analyte which diffuses into the sensor. Examples of usefulanticlotting agents include heparin and tissue plasminogen activator(TPA), as well as other known anticlotting agents.

The anticlotting agent may be applied to at least a portion of that partof the sensor that is to be implanted. The anticlotting agent may beapplied, for example, by bath, spraying, brushing, or dipping. Theanticlotting agent is allowed to dry on the sensor. The anticlottingagent may be immobilized on the surface of the sensor or it may beallowed to diffuse away from the sensor surface. Typically, thequantities of anticlotting agent disposed on the sensor are far belowthe amounts typically used for treatment of medical conditions involvingblood clots and, therefore, have only a limited, localized effect.

Insertion Device

An insertion device can be used to subcutaneously insert the sensor intothe patient. The insertion device is typically formed using structurallyrigid materials, such as metal or rigid plastic. Preferred materialsinclude stainless steel and ABS (acrylonitrile-butadiene-styrene)plastic. In some embodiments, the insertion device is pointed and/orsharp at the tip to facilitate penetration of the skin of the patient. Asharp, thin insertion device may reduce pain felt by the patient uponinsertion of the sensor. In other embodiments, the tip of the insertiondevice has other shapes, including a blunt or flat shape. Theseembodiments may be particularly useful when the insertion device doesnot penetrate the skin but rather serves as a structural support for thesensor as the sensor is pushed into the skin.

Sensor Control Unit

The sensor control unit can be integrated in the sensor, part or all ofwhich is subcutaneously implanted or it can be configured to be placedon the skin of a patient. The sensor control unit is optionally formedin a shape that is comfortable to the patient and which may permitconcealment, for example, under a patient's clothing. The thigh, leg,upper arm, shoulder, or abdomen are convenient parts of the patient'sbody for placement of the sensor control unit to maintain concealment.However, the sensor control unit may be positioned on other portions ofthe patient's body. One embodiment of the sensor control unit has athin, oval shape to enhance concealment. However, other shapes and sizesmay be used.

The particular profile, as well as the height, width, length, weight,and volume of the sensor control unit may vary and depends, at least inpart, on the components and associated functions included in the sensorcontrol unit. In general, the sensor control unit includes a housingtypically formed as a single integral unit that rests on the skin of thepatient. The housing typically contains most or all of the electroniccomponents of the sensor control unit.

The housing of the sensor control unit may be formed using a variety ofmaterials, including, for example, plastic and polymeric materials,particularly rigid thermoplastics and engineering thermoplastics.Suitable materials include, for example, polyvinyl chloride,polyethylene, polypropylene, polystyrene, ABS polymers, and copolymersthereof. The housing of the sensor control unit may be formed using avariety of techniques including, for example, injection molding,compression molding, casting, and other molding methods. Hollow orrecessed regions may be formed in the housing of the sensor controlunit. The electronic components of the sensor control unit and/or otheritems, such as a battery or a speaker for an audible alarm, may beplaced in the hollow or recessed areas.

The sensor control unit is typically attached to the skin of thepatient, for example, by adhering the sensor control unit directly tothe skin of the patient with an adhesive provided on at least a portionof the housing of the sensor control unit which contacts the skin or bysuturing the sensor control unit to the skin through suture openings inthe sensor control unit.

When positioned on the skin of a patient, the sensor and the electroniccomponents within the sensor control unit are coupled via conductivecontacts. The one or more working electrodes, counter electrode (orcounter/reference electrode), optional reference electrode, and optionaltemperature probe are attached to individual conductive contacts. Forexample, the conductive contacts are provided on the interior of thesensor control unit. Other embodiments of the sensor control unit havethe conductive contacts disposed on the exterior of the housing. Theplacement of the conductive contacts is such that they are in contactwith the contact pads on the sensor when the sensor is properlypositioned within the sensor control unit.

Sensor Control Unit Electronics

The sensor control unit also typically includes at least a portion ofthe electronic components that operate the sensor and the analytemonitoring device system. The electronic components of the sensorcontrol unit typically include a power supply for operating the sensorcontrol unit and the sensor, a sensor circuit for obtaining signals fromand operating the sensor, a measurement circuit that converts sensorsignals to a desired format, and a processing circuit that, at minimum,obtains signals from the sensor circuit and/or measurement circuit andprovides the signals to an optional transmitter. In some embodiments,the processing circuit may also partially or completely evaluate thesignals from the sensor and convey the resulting data to the optionaltransmitter and/or activate an optional alarm system if the analytelevel exceeds a threshold. The processing circuit often includes digitallogic circuitry.

The sensor control unit may optionally contain a transmitter fortransmitting the sensor signals or processed data from the processingcircuit to a receiver/display unit; a data storage unit for temporarilyor permanently storing data from the processing circuit; a temperatureprobe circuit for receiving signals from and operating a temperatureprobe; a reference voltage generator for providing a reference voltagefor comparison with sensor-generated signals; and/or a watchdog circuitthat monitors the operation of the electronic components in the sensorcontrol unit.

Moreover, the sensor control unit may also include digital and/or analogcomponents utilizing semiconductor devices, such as transistors. Tooperate these semiconductor devices, the sensor control unit may includeother components including, for example, a bias control generator tocorrectly bias analog and digital semiconductor devices, an oscillatorto provide a clock signal, and a digital logic and timing component toprovide timing signals and logic operations for the digital componentsof the circuit.

As an example of the operation of these components, the sensor circuitand the optional temperature probe circuit provide raw signals from thesensor to the measurement circuit. The measurement circuit converts theraw signals to a desired format, using for example, a current-to-voltageconverter, current-to-frequency converter, and/or a binary counter orother indicator that produces a signal proportional to the absolutevalue of the raw signal. This may be used, for example, to convert theraw signal to a format that can be used by digital logic circuits. Theprocessing circuit may then, optionally, evaluate the data and providecommands to operate the electronics.

Calibration

Sensors may be configured to require no system calibration or no usercalibration. For example, a sensor may be factory calibrated and neednot require further calibrating. In certain embodiments, calibration maybe required, but may be done without user intervention, i.e., may beautomatic. In those embodiments in which calibration by the user isrequired, the calibration may be according to a predetermined scheduleor may be dynamic, i.e., the time for which may be determined by thesystem on a real-time basis according to various factors, such as butnot limited to glucose concentration and/or temperature and/or rate ofchange of glucose, etc.

In addition to a transmitter, an optional receiver may be included inthe sensor control unit. In some cases, the transmitter is atransceiver, operating as both a transmitter and a receiver. Thereceiver may be used to receive calibration data for the sensor. Thecalibration data may be used by the processing circuit to correctsignals from the sensor. This calibration data may be transmitted by thereceiver/display unit or from some other source such as a control unitin a doctor's office. In addition, the optional receiver may be used toreceive a signal from the receiver/display units to direct thetransmitter, for example, to change frequencies or frequency bands, toactivate or deactivate the optional alarm system and/or to direct thetransmitter to transmit at a higher rate.

Calibration data may be obtained in a variety of ways. For instance, thecalibration data may simply be factory-determined calibrationmeasurements which can be input into the sensor control unit using thereceiver or may alternatively be stored in a calibration data storageunit within the sensor control unit itself (in which case a receiver maynot be needed). The calibration data storage unit may be, for example, areadable or readable/writeable memory circuit.

Calibration may be accomplished using an in vitro test strip (or otherreference), e.g., a small sample test strip such as a test strip thatrequires less than about 1 microliter of sample (for example Freestyle®blood glucose monitoring test strips from Abbott Diabetes Care). Forexample, test strips that require less than about 1 nanoliter of samplemay be used. In certain embodiments, a sensor may be calibrated usingonly one sample of body fluid per calibration event. For example, a userneed only lance a body part one time to obtain sample for a calibrationevent (e.g., for a test strip), or may lance more than one time within ashort period of time if an insufficient volume of sample is firstlyobtained. Embodiments include obtaining and using multiple samples ofbody fluid for a given calibration event, where glucose values of eachsample are substantially similar. Data obtained from a given calibrationevent may be used independently to calibrate or combined with dataobtained from previous calibration events, e.g., averaged includingweighted averaged, etc., to calibrate. In certain embodiments, a systemneed only be calibrated once by a user, where recalibration of thesystem is not required.

Alternative or additional calibration data may be provided based ontests performed by a doctor or some other professional or by thepatient. For example, it is common for diabetic individuals to determinetheir own blood glucose concentration using commercially availabletesting kits. The results of this test is input into the sensor controlunit either directly, if an appropriate input device (e.g., a keypad, anoptical signal receiver, or a port for connection to a keypad orcomputer) is incorporated in the sensor control unit, or indirectly byinputting the calibration data into the receiver/display unit andtransmitting the calibration data to the sensor control unit.

Other methods of independently determining analyte levels may also beused to obtain calibration data. This type of calibration data maysupplant or supplement factory-determined calibration values.

In some embodiments of the invention, calibration data may be requiredat periodic intervals, for example, every eight hours, once a day, oronce a week, to confirm that accurate analyte levels are being reported.Calibration may also be required each time a new sensor is implanted orif the sensor exceeds a threshold minimum or maximum value or if therate of change in the sensor signal exceeds a threshold value. In somecases, it may be necessary to wait a period of time after theimplantation of the sensor before calibrating to allow the sensor toachieve equilibrium. In some embodiments, the sensor is calibrated onlyafter it has been inserted. In other embodiments, no calibration of thesensor is needed.

Analyte Monitoring Device

In some embodiments of the invention, the analyte monitoring deviceincludes a sensor control unit and a sensor. In these embodiments, theprocessing circuit of the sensor control unit is able to determine alevel of the analyte and activate an alarm system if the analyte levelexceeds a threshold. The sensor control unit, in these embodiments, hasan alarm system and may also include a display, such as an LCD or LEDdisplay.

A threshold value is exceeded if the datapoint has a value that isbeyond the threshold value in a direction indicating a particularcondition. For example, a datapoint which correlates to a glucose levelof 200 mg/dL exceeds a threshold value for hyperglycemia of 180 mg/dL,because the datapoint indicates that the patient has entered ahyperglycemic state. As another example, a datapoint which correlates toa glucose level of 65 mg/dL exceeds a threshold value for hypoglycemiaof 70 mg/dL because the datapoint indicates that the patient ishypoglycemic as defined by the threshold value. However, a datapointwhich correlates to a glucose level of 75 mg/dL would not exceed thesame threshold value for hypoglycemia because the datapoint does notindicate that particular condition as defined by the chosen thresholdvalue.

An alarm may also be activated if the sensor readings indicate a valuethat is beyond a measurement range of the sensor. For glucose, thephysiologically relevant measurement range is typically about 50 to 250mg/dL, preferably about 40-300 mg/dL and ideally 30-400 mg/dL, ofglucose in the interstitial fluid.

The alarm system may also, or alternatively, be activated when the rateof change or acceleration of the rate of change in analyte levelincrease or decrease reaches or exceeds a threshold rate oracceleration. For example, in the case of a subcutaneous glucosemonitor, the alarm system might be activated if the rate of change inglucose concentration exceeds a threshold value which might indicatethat a hyperglycemic or hypoglycemic condition is likely to occur.

A system may also include system alarms that notify a user of systeminformation such as battery condition, calibration, sensor dislodgment,sensor malfunction, etc. Alarms may be, for example, auditory and/orvisual. Other sensory-stimulating alarm systems may be used includingalarm systems which heat, cool, vibrate, or produce a mild electricalshock when activated.

Drug Delivery System

The subject invention also includes sensors used in sensor-based drugdelivery systems. The system may provide a drug to counteract the highor low level of the analyte in response to the signals from one or moresensors. Alternatively, the system may monitor the drug concentration toensure that the drug remains within a desired therapeutic range. Thedrug delivery system may include one or more (e.g., two or more)sensors, a processing unit such as a transmitter, a receiver/displayunit, and a drug administration system. In some cases, some or allcomponents may be integrated in a single unit. A sensor-based drugdelivery system may use data from the one or more sensors to providenecessary input for a control algorithm/mechanism to adjust theadministration of drugs, e.g., automatically or semi-automatically. Asan example, a glucose sensor may be used to control and adjust theadministration of insulin from an external or implanted insulin pump.

EXAMPLES

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the present invention, and are not intended to limit thescope of what the inventors regard as their invention nor are theyintended to represent that the experiments below are all or the onlyexperiments performed. Efforts have been made to ensure accuracy withrespect to numbers used (e.g. amounts, temperature, etc.) but someexperimental errors and deviations should be accounted for. Unlessindicated otherwise, parts are parts by weight, molecular weight isweight average molecular weight, temperature is in degrees Centigrade,and pressure is at or near atmospheric.

Example 1 Reference Electrode Having a Dielectric Cover Layer

FIG. 6A shows a glucose sensor design with three electrodes that areformed by screen-printing on a polyester substrate. The conductivetraces are separated by dielectric layers that are also formed by screenprinting. The Ag/AgCl pad has an area of about 0.1 mm², and a thicknessof about 5 μm. The total load of AgCl is about 400 ng. Clinical studiesusing sensors of this design have shown that the sensor output currentbecomes noisy 3-4 days after implantation. Subsequent in-vitro studiesconfirmed this phenomenon. Coulometric experiments showed that, whenevera sensor's output current became noisy, there was almost no AgCl left onits Ag/AgCl reference electrode.

To improve the sensor performance, a series of sensors were fabricatedwith the Ag/AgCl pad covered partially by a layer of dielectric. Threesets of sensors with exposed Ag/AgCl areas of approximately 20%, 50%,and 100% (n=4 for each case) of the relatively constant total electrodearea were tested in vitro. The results are illustrated in FIG. 7. It canbe seen that the sensor life overall was greatly prolonged by covering aportion of the Ag/AgCl of the reference electrode with a dielectric.

Based on this result, a new sensor design with a one-layer lessstructure is shown schematically in FIG. 6B. By using only the Ag/AgCltrace, this design not only eliminates one screen-printing step, butalso ensures the consistency of reference electrode layout.

Example 2 Reference Electrode Having a Permselective Layer

A permselective coating (in addition to the overlying glucoseflux-limiting membrane) can extend reference electrode lifetime.Experiments were performed with a localized coating ofpoly(vinylpyridine-co-styrene) with 10% loading of styrene over theAg/AgCl reference element of the reference electrode. This material hasa much lower permeability than the standard (control) glucoseflux-limiting membrane. FIG. 8 shows accelerated aging studies (at 66°C.) of such an electrode, compared to a control (non-protected)reference electrode.

Note electrode lifetime (the time until onset of noise in the signal) issignificantly improved in the protected electrode, from approximately 18hours to 120 hours, or about a 6 fold increase. Furthermore, thisprotected electrode is expected to have a substantially longer lifetimeat the subcutaneous temperature of about 34° C.

Example 3 Renewing Ag/AgCl Reference Electrode In Situ with an AppliedElectrical Potential

The reference electrode of the experimental sensors was formed by screenprinting a Ag/AgCl pad with an area of about 0.1 mm², and a thickness ofabout 5 μm. The total load of AgCl and Ag are about 400 ng and 1000 ng,respectively. The total electrochemically accessible AgCl and Ag areequivalent to 200 μC and 500 μC, respectively. In order to evaluate thecharge injection frequency, the dissolution rate of AgCl was firstdetermined in-vitro in PBS using standard coulometric technique. As seenin FIG. 9, AgCl dissolves at an average rate of about 50 μC per day. Thevariation in the charge values is due to the variation of the referenceelectrode pad areas caused by the screen printing resolution limitation.

With a potential of +200 mV against a commercial standard Ag/AgClelectrode (Bioanalytical Systems, Inc.), AgCl was successfullyregenerated repeatedly on the sensor's reference electrode, asdemonstrated in FIGS. 10A and 10B. In addition, it can be seen that theloss rate of the regenerated AgCl is about the same as the original AgClfrom the screen printing ink.

In order to regenerate AgCl in-situ, this invention utilizes the workingelectrode of a Navigator sensor as the reference electrode. Therationale is that the Os(III/II) redox couple present in the sensingchemistry should dominate the electrochemical potential of theelectrode. In order to select a proper potential to regenerate AgClusing this electrode as a reference, the potential range of theOs(III/II) couple on the sensor's working electrode was first determinedGlucose and oxygen concentrations are the two variables in the in-vivoenvironment affecting the relative ratio of Os(III) and Os(II) whichdetermines the electrode potential (per Nernst equation). High glucoseand low oxygen concentrations would raise the Os(II) relative toOs(III), and this would shift the electrode potential negatively. It wasfound that the possible potential range of the Os couple against astandard Ag/AgCl is from 0 to −250 mV when varying glucose from 0 to 30mM and O₂ from 20% to 2%. As shown in FIG. 11, a potential higher than+450 mV vs. Os(III/II) would be equivalent to +200 mV or higher vs. astandard Ag/AgCl electrode.

Based on this result, a potential of +500 mV vs. the working electrodewas selected to regenerate AgCl on the sensor' reference electrodein-situ. As shown in FIG. 12, the results are the same as using externalelectrodes as shown in FIGS. 10A-10B. In both situations a stabilizationof the signal was witnessed after the level of AgCl was replenished byapplication of electrical potential.

The preceding merely illustrates the principles of the invention. Itwill be appreciated that those skilled in the art will be able to devisevarious arrangements which, although not explicitly described or shownherein, embody the principles of the invention and are included withinits spirit and scope. Furthermore, all examples and conditional languagerecited herein are principally intended to aid the reader inunderstanding the principles of the invention and the conceptscontributed by the inventors to furthering the art, and are to beconstrued as being without limitation to such specifically recitedexamples and conditions. Moreover, all statements herein recitingprinciples, aspects, and embodiments of the invention as well asspecific examples thereof, are intended to encompass both structural andfunctional equivalents thereof. Additionally, it is intended that suchequivalents include both currently known equivalents and equivalentsdeveloped in the future, i.e., any elements developed that perform thesame function, regardless of structure. The scope of the presentinvention, therefore, is not intended to be limited to the exemplaryembodiments shown and described herein. Rather, the scope and spirit ofpresent invention is embodied by the appended claims.

That which is claimed is:
 1. A method for monitoring a level of ananalyte, the method comprising: inserting an electrochemical sensor intoskin of a patient, wherein the electrochemical sensor comprises: aworking electrode comprising a sensing layer disposed thereon; and areference electrode comprising a layer of Ag/AgCl having a width that isequal to the width of the working electrode and a layer of dielectricmaterial having a width that is at least equal to the width of the layerof Ag/AgCl and a length that is at least equal to the length of thelayer of Ag/AgCl disposed on top of the layer of Ag/AgCl; collectingdata regarding a level of an analyte from signals generated by thesensor; transmitting the collected data to a display unit; anddisplaying an indication of the level of the analyte on the display ofthe display unit.
 2. The method of claim 1, wherein the analyte isglucose.
 3. The method of claim 1, wherein collecting data comprisesgenerating signals from the sensor and processing the signals into data.4. The method of claim 1, wherein the data comprises the signals fromthe sensor.
 5. The method of claim 1, further comprising activating analarm if the data indicates an alarm condition.
 6. The method of claim1, further comprising administering a drug in response to the data. 7.The method of claim 6, wherein the drug is insulin.
 8. The method ofclaim 7, further comprising obtaining a calibration value from acalibration device to calibrate the data.
 9. The method of claim 8,wherein the calibration device is coupled to the display unit.
 10. Themethod of claim 9, further comprising transmitting the calibration valuefrom a transmitter in the display unit to a receiver in the sensorcontrol unit.
 11. The method of claim 1, wherein the sensing layer ofthe working electrode comprises a glucose-responsive enzyme.
 12. Themethod of claim 1, wherein the sensing layer of the working electrodecomprises a redox mediator.
 13. The method of claim 12, wherein theredox mediator comprises a complex selected from the group consisting ofa ruthenium-containing complex and an osmium-containing complex.
 14. Themethod of claim 12, wherein the redox mediator is non-leachable withrespect to the working electrode.
 15. The method of claim 12, whereinthe redox mediator is immobilized on the working electrode.
 16. Themethod of claim 1, further comprising a flux limiting membrane disposedover the sensing layer on the working electrode.
 17. The method of claim1, further comprising a second dielectric layer disposed on at least aportion of the working electrode.
 18. The method of claim 17, whereinthe reference electrode is disposed over the second dielectric layer.19. The method of claim 18, further comprising a counter electrodedisposed on top of the first dielectric layer and a third dielectriclayer disposed on top of the counter electrode.